PA1-931

antibody from Invitrogen Antibodies

Targeting: CALB1 CALB

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-931

Provider

Invitrogen Antibodies

Product name

Calbindin D28K Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Purifed from natural sources

Description

PA1-931 detects calbindin D-28k protein in rat samples. PA1-931 has successfully been used in Western blot , ELISA, immunoprecipitation and immunohistochemical procedures. By Western blot, this antibody detects a 28 kDa protein representing calbindin D from rat cerebellum and rat kidney extract. Immunohistochemical staining on paraffin sections demonstrates that calbindin D is localized to the cytoplasm in epithelial cells of distal tubules in human kidney cortex after staining with PA1-931. The PA1-931 immunizing protein corresponds to the purified 28 kDa calbindin-D protein from rat kidney. Reconstitute in 100 µL PBS to create a stock of 1 mg/mL.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Conditional Deletion of Activating Rearranged During Transfection Receptor Tyrosine Kinase Leads to Impairment of Photoreceptor Ribbon Synapses and Disrupted Visual Function in Mice.

Peng WH, Liao ML, Huang WC, Liu PK, Levi SR, Tseng YJ, Lee CY, Yeh LK, Chen KJ, Chien CL, Wang NK
Frontiers in neuroscience 2021;15:728905

---

Lycii radicis cortex inhibits glucocorticoid‑induced bone loss by downregulating Runx2 and BMP‑2 expression.

Lee B, Hong S, Kim M, Kim EY, Park HJ, Jung HS, Kim JH, Sohn Y
International journal of molecular medicine 2021 Aug;48(2)

---

Anti-Yo antibody uptake and interaction with its intracellular target antigen causes Purkinje cell death in rat cerebellar slice cultures: a possible mechanism for paraneoplastic cerebellar degeneration in humans with gynecological or breast cancers.

Greenlee JE, Clawson SA, Hill KE, Wood B, Clardy SL, Tsunoda I, Carlson NG
PloS one 2015;10(4):e0123446

---

Developmental and age-dependent changes of 28-kDa calbindin-D in the central nervous tissue determined with a sensitive immunoassay method.

Kurobe N, Inaguma Y, Shinohara H, Semba R, Inagaki T, Kato K
Journal of neurochemistry 1992 Jan;58(1):128-34

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of calbindin D from rat kidney using Product # PA1-931.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed using whole cell extracts (30 µg) of Mouse Brain (Lane 1), Rat Brain (Lane 2), IMR-32 (Lane 3), SH-SY5Y (Lane 4) and U-2 OS (Lane 5).The blots were probed with Anti-Calbindin D-28K Rabbit Polyclonal Antibody (Product # PA1-931, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4µg/mL 1:2500 dilution). A ~28 kDa band corresponding to Calbindin D-28K was observed across tissues and cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockout of Calbindin D28K was achieved by CRISPR-Cas9 genome editing. Western blot analysis of Calbindin D28K was performed by loading 20 µg of SH-SY5Y wild type (Lane 1), SH-SY5Y Cas9 control (Lane 2), SH-SY5Y Calbindin D28K knockout (Lane 3) whole cell extracts. The blot was probed with Anti-Calbindin D28K Polyclonal Antibody (Product # PA1-931) using 2 µg/mL and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036) using 1:4000 dilution. Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to Calbindin D28K. Uncharacterised bands were observed in WT and Cas9 samples at ~170 kDa.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig 4 Comparison of uptake and cytotoxicity of anti-Yo antibody versus three other antibodies specific for the intracellular Purkinje cell proteins: calbindin, calmodulin, and PCP-2. The top row of panels demonstrates uptake and accumulation of IgG (red) within Purkinje cells after 96 hours in cultures incubated with anti-calmodulin, anti-calbindin, anti PCP-2, or anti-Yo IgGs. Entry of SYTOX green into Purkinje cells containing IgG, indicative of cell membrane injury and death (yellow), was seen in only in cultures incubated with anti-Yo IgGs (examples shown by arrows). The lower panels show only SYTOX green staining of Purkinje cells in cultures incubated with the antibodies indicated. In the culture incubated with anti-PCP-2. SYTOX staining indicative of cell death is seen in a single cell outside the Purkinje cell layer (asterisk) but not in Purkinje cells. Cultures were followed through 144 hours. There was progression of cell death in cultures incubated with anti-Yo antibodies. In contrast, cultures incubated calbindin, calmodulin, and PCP-2 did not exhibit Purkinje cell death above background levels seen in controls incubated with normal human IgG (data not shown). Scale bar = 20mu.