Gibco heat stable recombinant human bFGF

Engineered for greater stability

Native bFGF is highly unstable under normal cell culture conditions with a half-life of <8 hours. Gibco Heat Stable Recombinant Human bFGF has been engineered for greater stability in cell culture conditions, sustaining ≥ 80% activity at 37°C for at least 72 hours.

  • Superior performance: retains bioactivity without the use of artificially high concentrations
  • Easy to use: direct replacement for your current bFGF allowing for easy substitution in protocols
  • Advancement without compromise: >90% sequence homology to native bFGF, leaving heparin and FGF receptor binding sites untouched

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Try it today: Free Heat Stable bFGF samples

Try Gibco Heat Stable Recombinant Human bFGF for greater stability and improved consistency in your cell cultures.

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Superior cell culture performance

HS bFGF maintains bFGF activity at 37°C

graph of proliferation vs dose for HS bFGF
graph of proliferation vs dose for native bFGF
Figure 1. Cell proliferation analysis studies demonstrate HS bFGF exhibits superior activity compared to native bFGF. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The 10 ng/mL HS bFGF (left) and native bFGF (right) solutions were stored at 4°C (unstressed) or 37°C (heat stressed) for 72 hours before being used to treat the Balb/3T3 cells in a dose-dependent manner.

Over 80% sustained biological activity

bar chart of proliferation for HS bFGF and native bFGF
Figure 2. HS bFGF demonstrates greater activity after 72 hours of heat stress than the native protein. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The percent activity is amount of activity maintained after heat stress, relative to the same solution stored at 4°C (unstressed). The 10 ng/mL HS bFGF and native bFGF solutions were stored at 4°C or 37°C for 72 hours before being used to treat the Balb/3T3 cells.

HS bFGF maintains neural stem cell multipotency

phase-contrast and immunostained images of NSCs cultured in Gibco HS bFGF
Figure 3. Heat Stable bFGF maintains the morphology and SOX1 expression of multipotent NSCs. Phase contrast (left) and immunostained (right) photomicrographs of NSCs cultured in the presence of HS bFGF (top row) and native bFGF (bottom row). Immunostaining for NSC multipotency marker SOX1 (red) with nuclear (DAPI) counterstain (blue).

HS bFGF does not interfere with undirected differentiation

immunostained images of NSCs cultured in Gibco HS bFGF showing expected differentiation markers

Figure 4. Spontaneous differentiation supported by HS bFGF. Primary rat NSC isolated from Sprague Dawley E14 cortex cultured for 3 passages in DMEM/F12 + GlutaMAX supplemented with N2, NEAA, β-mercaptoethanol and 10 ng/mL HS bFGF. Media changes made every 48 hours without daily spiking of bFGF. At passage 3, cells were allowed to spontaneously differentiate by culturing in growth medium without HS bFGF. Cells showed equivalent potential to undergo undirected differentiation as compared to native bFGF, measured by markers MAP2 (neuron), GFAP (astrocytes) and GALC (oligodendrocytes).

HS bFGF enhances growth of cancer spheroids via a more streamlined protocol

brightfield microscopy image of cancer spheroids grown with Gibco HS bFGF

Figure 5. HS bFGF enhances growth of cancer spheroids without the need for troublesome media changes. Spheroids were formed in Nunclon Sphera microplates in serum-free medium containing no bFGF, 10 ng/mL native bFGF, or 10 ng/mL HS bFGF. After 8 days in culture without media changes, the cells were stained via live (green)-dead (red) assay and DAPI counterstain (blue). All spheroids were viable, but the spheroid exposed to HS bFGF was the largest of the three experimental conditions. For more information please see our application note

Gene expression of cells treated with HS bFGF is comparable to that of cells treated with native bFGF

gene expression heatmap showing correlation between the two treatments
 Click image to enlarge
Figure 6. bFGF-target and off-target gene expression was comparable in cells treated with native bFGF and Heat Stable bFGF. The TaqMan Array Human Signal Transduction Pathways array (Cat. No. 4418775) and the TaqMan Array Human FGF Pathway (Cat. No. 4414136) were used to assess expression of genes in the androgen, calcium, CREB, estrogen, hedgehog, insulin, JAK-STAT, LDL, mitogenic, NFAT, NFkB, p53, phospholipase C, protein kinase C, retinoic acid, stress, survival, TGF-β, Wnt, and FGF-associated pathways, respectively. Expression was assessed using human induced pluripotent stem cells treated with 10 ng/mL native bFGF or Heat Stable bFGF. Genes from both arrays were separated into ‘bFGF-target’ or ‘off-target’ based on their downstream relationship to bFGF, as is currently indicated by the literature. The bFGF-target and off-target panels show the results from 43 and 71 genes, respectively.

HS bFGF outperforms top competitors

bar chart of bFGF activity from Gibco HS bFGF and three competitor products

Figure 7. HS bFGF exhibits superior activity after extended 37°C incubation compared to competitor’s products. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The percent activity is amount of activity maintained after heat stress (37°C), relative to the same solution stored at 4°C (unstressed). The 10 ng/mL HS bFGF and competitor bFGF solutions were stored at 4°C or 37°C for 72 hours before being used to treat the Balb/3T3 cells, N≥3. Mean ± SEM.

HS bFGF facilitates stem cell culture workflows

4-panel microscopy image showing iPSCs cultured with Gibco HS bFGF
Figure 8. Comparison iPSCs cultured in the presence of Gibco HS bFGF or StemBeads reagent. Microscopic images of StemBeads reagent (10 ng/mL) captured at (A) 10x and (B) 20x magnification. Because StemBeads beads vary in size (arrows indicate different sized StemBeads particles), they can be difficult to distinguish from cells and also from debris. Twenty-four hours after seeding human iPSCs in the presence of either (C) 10 ng/mL Gibco HS bFGF or (D) 10 ng/mL StemBeads reagent, phase-contrast images were captured of the cultured cells. The image of cells cultured with HS bFGF is clear, containing minimal debris, whereas the StemBeads reagent–containing culture shows more apparent debris. In addition because the beads tend to sink relatively quickly in solution, accurately measuring StemBeads reagent concentration can present a challenge.

HS bFGF maintains bFGF activity at 37°C

graph of proliferation vs dose for HS bFGF
graph of proliferation vs dose for native bFGF
Figure 1. Cell proliferation analysis studies demonstrate HS bFGF exhibits superior activity compared to native bFGF. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The 10 ng/mL HS bFGF (left) and native bFGF (right) solutions were stored at 4°C (unstressed) or 37°C (heat stressed) for 72 hours before being used to treat the Balb/3T3 cells in a dose-dependent manner.

Over 80% sustained biological activity

bar chart of proliferation for HS bFGF and native bFGF
Figure 2. HS bFGF demonstrates greater activity after 72 hours of heat stress than the native protein. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The percent activity is amount of activity maintained after heat stress, relative to the same solution stored at 4°C (unstressed). The 10 ng/mL HS bFGF and native bFGF solutions were stored at 4°C or 37°C for 72 hours before being used to treat the Balb/3T3 cells.

HS bFGF maintains neural stem cell multipotency

phase-contrast and immunostained images of NSCs cultured in Gibco HS bFGF
Figure 3. Heat Stable bFGF maintains the morphology and SOX1 expression of multipotent NSCs. Phase contrast (left) and immunostained (right) photomicrographs of NSCs cultured in the presence of HS bFGF (top row) and native bFGF (bottom row). Immunostaining for NSC multipotency marker SOX1 (red) with nuclear (DAPI) counterstain (blue).

HS bFGF does not interfere with undirected differentiation

immunostained images of NSCs cultured in Gibco HS bFGF showing expected differentiation markers

Figure 4. Spontaneous differentiation supported by HS bFGF. Primary rat NSC isolated from Sprague Dawley E14 cortex cultured for 3 passages in DMEM/F12 + GlutaMAX supplemented with N2, NEAA, β-mercaptoethanol and 10 ng/mL HS bFGF. Media changes made every 48 hours without daily spiking of bFGF. At passage 3, cells were allowed to spontaneously differentiate by culturing in growth medium without HS bFGF. Cells showed equivalent potential to undergo undirected differentiation as compared to native bFGF, measured by markers MAP2 (neuron), GFAP (astrocytes) and GALC (oligodendrocytes).

HS bFGF enhances growth of cancer spheroids via a more streamlined protocol

brightfield microscopy image of cancer spheroids grown with Gibco HS bFGF

Figure 5. HS bFGF enhances growth of cancer spheroids without the need for troublesome media changes. Spheroids were formed in Nunclon Sphera microplates in serum-free medium containing no bFGF, 10 ng/mL native bFGF, or 10 ng/mL HS bFGF. After 8 days in culture without media changes, the cells were stained via live (green)-dead (red) assay and DAPI counterstain (blue). All spheroids were viable, but the spheroid exposed to HS bFGF was the largest of the three experimental conditions. For more information please see our application note

Gene expression of cells treated with HS bFGF is comparable to that of cells treated with native bFGF

gene expression heatmap showing correlation between the two treatments
 Click image to enlarge
Figure 6. bFGF-target and off-target gene expression was comparable in cells treated with native bFGF and Heat Stable bFGF. The TaqMan Array Human Signal Transduction Pathways array (Cat. No. 4418775) and the TaqMan Array Human FGF Pathway (Cat. No. 4414136) were used to assess expression of genes in the androgen, calcium, CREB, estrogen, hedgehog, insulin, JAK-STAT, LDL, mitogenic, NFAT, NFkB, p53, phospholipase C, protein kinase C, retinoic acid, stress, survival, TGF-β, Wnt, and FGF-associated pathways, respectively. Expression was assessed using human induced pluripotent stem cells treated with 10 ng/mL native bFGF or Heat Stable bFGF. Genes from both arrays were separated into ‘bFGF-target’ or ‘off-target’ based on their downstream relationship to bFGF, as is currently indicated by the literature. The bFGF-target and off-target panels show the results from 43 and 71 genes, respectively.

HS bFGF outperforms top competitors

bar chart of bFGF activity from Gibco HS bFGF and three competitor products

Figure 7. HS bFGF exhibits superior activity after extended 37°C incubation compared to competitor’s products. Activity was measured with PrestoBlue dye analysis of Balb/3T3 cell proliferation. The percent activity is amount of activity maintained after heat stress (37°C), relative to the same solution stored at 4°C (unstressed). The 10 ng/mL HS bFGF and competitor bFGF solutions were stored at 4°C or 37°C for 72 hours before being used to treat the Balb/3T3 cells, N≥3. Mean ± SEM.

HS bFGF facilitates stem cell culture workflows

4-panel microscopy image showing iPSCs cultured with Gibco HS bFGF
Figure 8. Comparison iPSCs cultured in the presence of Gibco HS bFGF or StemBeads reagent. Microscopic images of StemBeads reagent (10 ng/mL) captured at (A) 10x and (B) 20x magnification. Because StemBeads beads vary in size (arrows indicate different sized StemBeads particles), they can be difficult to distinguish from cells and also from debris. Twenty-four hours after seeding human iPSCs in the presence of either (C) 10 ng/mL Gibco HS bFGF or (D) 10 ng/mL StemBeads reagent, phase-contrast images were captured of the cultured cells. The image of cells cultured with HS bFGF is clear, containing minimal debris, whereas the StemBeads reagent–containing culture shows more apparent debris. In addition because the beads tend to sink relatively quickly in solution, accurately measuring StemBeads reagent concentration can present a challenge.

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FAQs

Question Answer
What are the possible abbreviations for basic fibroblast growth factor? Basic fibroblast growth factor can be notated in the following ways: bFGF, FGF2, FGF-2, FGF-β, FGF-basic.
What is heat stable? Heat stable is the ability to sustain activity at elevated temperatures (37°C). Also known as thermostable.
What kind of tubes should be used to store aliquots? Polypropylene tubes should be used to store aliquots of reconstituted Heat Stable Recombinant Human bFGF.
Does the His-tag on Heat Stable Recombinant Human bFGF affect biological activity? No, the His-tag has no effect on biological activity.
Is Heat Stable Recombinant Human bFGF full length? Yes, Heat Stable Recombinant Human bFGF is full-length at 155 amino acids, plus 20 amino acids for N-terminal tag for purification purposes.
How do I determine a working concentration? We recommend that you use the same working concentration as native human bFGF that you are currently using. Cell cultures should be monitored to determine whether the working concentration can be reduced.
Can Heat Stable Recombinant Human bFGF be used in place of native recombinant human bFGF? Yes, Heat Stable Recombinant Human bFGF can be used in place of native human bFGF. It is a direct replacement, offering greater stability in cell culture conditions.
Was any additive added to achieve thermostability of Heat Stable Recombinant Human bFGF? No additives were used. Heat Stable Recombinant Human bFGF was engineered to maintain bioactivity at 37°C. The patent pending technology has >90% sequence homology to native human bFGF.
Has protein engineering altered the activity of Heat Stable Recombinant Human bFGF? HS bFGF does not exhibit higher bioactivity, nor does it stimulate cell signaling differently than native bFGF.
Why should Heat Stable Recombinant Human bFGF be used instead of native recombinant human bFGF? Native human bFGF degrades at standard culture conditions (i.e., 37°C). This means that more frequent media changes and/or more protein is required to maintain biological activity. Heat Stable Recombinant Human bFGF retains bioactivity minimizing fluctuations to more closely mimic physiological conditions for cells.
How long can I use media supplemented with Heat Stable Recombinant Human bFGF? We recommend that you use growth factor-supplemented medium within 2-4 weeks of supplementation.
Can I freeze aliquots after reconstitution? Yes, aliquots can be frozen and stored at -20°C for up to 1 year from date of receipt. Avoid additional freeze/thaw cycles.
I am not able to resuspend the 5 μg size at the recommended concentration. How should I use it? The 5 μg Heat Stable Recombinant Human bFGF should be reconstituted directly in the intended culture medium, preferably media containing BSA or similar protein.

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